Analysis of Protein Posttranslational Modifications Using DIGE-Based Proteomics
Difference gel electrophoresis (DIGE) is most often used to assess relative changes in the expression levels of individual proteins in multiple complex samples, and this information is valuable in making inferences about relative protein activity. However, a protein’s activity is not solely dependent upon its expression level. A change in activity may also be influenced by myriad posttranslational modifications (PTMs), including palmitoylation, ubiquitination, oxidation, and phosphorylation. In this chapter, we describe the use of DIGE to determine specific PTMs by introducing specific labels or changes in pI and/or molecular weight.
Key wordsDifference gel electrophoresis Neuroproteomics Oxidation Palmitoylation Phosphory-lation Posttranslational modification Ubiquitination
This work was supported by the UNC Systems-Proteomics Center and the Duke Neuroproteomics Center.
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